摘要
目的:通过siRNA干扰技术沉默ARTN基因在人食管癌细胞TE11中的表达,研究其对人食管癌细胞增殖、迁移和侵袭能力的影响。方法:设计并合成3条特异性针对ARTN基因的siRNA,构建了真核表达载体pSilencer2.1-ARTN-siRNA,转染到TE11细胞中,经Real-time PCR和Western blot检测ARTN在mRNA和蛋白水平的表达;通过MTT比色法、细胞划痕实验和Boyden小室侵袭实验观察转染pSilencer2.1-ARTN-siRNA后细胞增殖、迁移和侵袭能力的改变。结果:转染重组质粒pSilencer2.1-ARTN-siRNA后人食管癌细胞TE11中ARTN的mRNA表达受到抑制,蛋白表达降低;通过体外实验表明,细胞的增殖能力、迁移能力和侵袭能力均减弱。结论:siRNA沉默ARTN基因的表达可降低人食管癌细胞TE11的增殖、迁移和侵袭能力,ARTN可能是食管癌的一个生物治疗靶点,有望成为治疗食管癌的新途径。
Objective： To investigate whether ARTN protein can affect proliferation, migration and invasion of esophageal carcinoma cell line TEll using small interfering IRNA. Methods： Three pairs of small interfering RNA （siRNA） specific to ARTN mRNA were designed and cloned into plasmid pSilencer2.1-U6-neo. The recombined eukaryotic expression vector pSilencer2.1-ARTN-siRNA was then transfected into TEll cells. The levels of ARTN mRNA and protein expression were detected using real-time PCR and Western blotting. The proliferation, migration and invasion of TE11-pSilencer2.1 cells and TE11-pSflencer2.1-ARTN-siRNA cells were assessed by MTT, wound healing and Boyden chamber. Results： The recombined eukaryotic expression vector pSilencer2.1-ARTN-siRNA was transfected successfully. After transfection, the levels of expression of ARTN mRNA and protein were significantly reduced in TEll- pSilencer2.1-siRNA-ARTN cells. The proliferation, migration and invasion of TE11-pSilencer2.1-siRNA-ARTN cells were also significantly decreased. Conclusion： Silencing the ARTN gene using siRNA can effectively suppress proliferation, migration and invasion of TEll esophageal carcinoma cells. ARTN may be a potential therapeutic target, and silencing the ARTN gene using siRNA may provide a novel therapeutic approach for esophageal cancer.
出处
《中国肿瘤临床》
CAS
CSCD
北大核心
2010年第24期1432-1436,共5页
Chinese Journal of Clinical Oncology
关键词
SIRNA
RTN
食管癌
侵袭
真核表达载体
Small interfering RNA
ARTN
Esophageal neoplasm
Invasiveness
Recombined eukaryotic expression vector
